CN1210394C - Natural inductive non bean family plant tumour forming nitrogen fixation technical method and bacterial spawn - Google Patents
Natural inductive non bean family plant tumour forming nitrogen fixation technical method and bacterial spawn Download PDFInfo
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- Fertilizers (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to a technical method and strains for nodulation and nitrogen fixation of natural inductive non-leguminous plants. A group of microbial floras are artificially bred from the nature by the present invention; the present invention can enable leguminous plants and more than 130 kinds of non-leguminous plants which can not carry out nodulation and nitrogen fixation in the root system originally to together carry out nodulation and nitrogen fixation. Under the condition of according with the nature as much as possible, namely 'open fermentation technology', the beneficial flora is industrially produced in a large scale to be prepared into broad-spectrum liquid biological material which can be used for various plants (comprising fungi-mushrooms); subsequently, under natural conditions, the solid fermentation process is carried out many times to produce solid biological fertilizer, which has important scientific significance and significant economic value for agricultural production necessary for the living of humans. The present invention can solve various problems of soil hardening, pollution to water resources and environment, damage to crop heritability, reduction of the quality of agricultural products, etc.
Description
Technical field
The present invention relates to a kind of spontaneous induction non-leguminous plant nodulation and nitrogen fixation technological method and bacterial classification.
Background technology
Some microorganism of nature can be an ammonia with airborne nitrogen transformation, be plant, animal and other microorganism growth provide nitrogenous source, be referred to as biological nitrogen fixation, according to Food and Argriculture OrganizationFAO (FAO) nineteen ninety-five guestimate, global every year is by nearly 200,000,000 tons of the nitrogen amount of biological fixation (being equivalent to 400,000,000 tons of urea), the scientific research in more than 100 year, recognize biological nitrogen fixation only provide can with the higher plant of nitrogen-fixing microorganism symbiosis dross, comprise leguminous plants and part non-leguminous plant (mainly being trees), the required nitrogen of growing, and other plant, particularly important food crop, fruit tree, vegetables etc., it is then limited that but symbiotic nitrogen-fixing microorganism with it not, the spontaneous nitrogen-fixing microorganism in the soil offer their nitrogen.Therefore, human increase, the high yield of required these crops just have to rely on chemical fertilizer.And, causing pollution, the variation of farm crop heritability of soil compaction, water system and environment because using chemical fertilizer adds chemical pesticide in a large number, quality of agricultural product descends and causes significant damage to the mankind again.
The mankind recognize that finally the development of the future of agriculture will turn to Organic farming once more from inorganic agriculture, and must expand non-leguminous plant to from leguminous plants for the research range of biological nitrogen fixation, the social needs that the technology of the present invention that Here it is produces.
Human research to biological nitrogen fixation in more than 100 year once produced many achievements such as leguminous crop Inoculant, chemical simulation fixed nitrogen, but limited to the effect of agriculture production; The graceful new manual-induced non-leguminous plant nodulation and nitrogen fixation of the method technology success of having announced of king in 1981 with mangneto mutagenesis, cause the attention of domestic and international agricultural and microbial world, but the expert authority who is not understood non-leguminous plant nodulation and nitrogen fixation technology soon negates to have become an academicly controversial intricate case without a clue., the Nie Yuan of Shandong Normal University was rich afterwards repeats non-leguminous plant dross and so-called 7 R﹠D institutions of quilt with the 2-4D hormone.But, therefore end up with nothing definite because its nitrogen fixation is not obvious.
Summary of the invention
The objective of the invention is for a kind of brand-new environmental friend fertilizer is provided, make farm output increase quality improving; After Wang Manxin, actual through the scientific researches in 17 years and the production of combining closely, studied successfully a kind of spontaneous induction non-leguminous plant nodulation and nitrogen fixation technological method and bacterial classification and many-sided utilisation technology method.
The characteristics of microorganism maximum are exactly very easily to make a variation.An effective fixed nitrogen bacterial classification is isolated in attempt in the laboratory, the problem that solves biological nitrogen fixation is impossible.Moreover be subjected to leguminous plants one non-leguminous plant, have growing nitrogen-fixing altogether from growing nitrogen-fixing, the limitation of autotrophic microorganism one these original theories of heterotrophic microorganism as not freeing from these mistaken ideas, solve the problem of " non-leguminous plant " nodulation and nitrogen fixation, and is impossible especially.Japan microorganism scholar proposes " agricultural microorganism research only goes back to nature, and is only the only way ", and this viewpoint is right as rain.Accepted by Canada, Germany and Australian many scholars.
The present invention gathers Rhizobium leguminosarum, non-pulse family root endophyte from nature just like this, through reasonable combination, enlarge under the condition near nature as far as possible, allow probiotics preponderate artificially, return in the physical environment, between the root system of soil and plant, mutagenesis in the whole process of plant-growth makes it to become one group of effective useful again combination flora, up to the effect that reaches non-leguminous plant root system nodulation and nitrogen fixation, and rejuvenation under field conditions (factors), prevent spawn degeneration.
Bacterial classification of the present invention, its sample is preserved at China Committee for Culture Collection of Microorganisms common micro-organisms center (hereinafter to be referred as the preservation center), preservation date: on June 12nd, 2003, its address is a No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City Institute of Microorganism, Academia Sinica, postcode: 100080, the bacterial classification of non-leguminous plant nodulation and nitrogen fixation is the combination flora, registers on the books to number and be in the bacterial classification of combination flora and preservation center thereof:
Register on the books and number 0949 in viride (Trichoderma viride) preservation center
Register on the books and number 0948 in root nodule bacterium (Rhizobium sp.) preservation center
Register on the books and number 0947 in look brown streptomycete (Streptomyces chromofuscus) preservation center
Register on the books and number 0946 in Bacillus foecalis alkaligenes (Alcaligenes faecalis) preservation center
Register on the books and number 0945 in edaphic bacillus (Agrobacterium sp.) preservation center
Register on the books and number 0944 in bacillus megaterium (Bacillus megateriium) preservation center
More than these bacterial classifications must combine, in 0 ℃ of preservation of low temperature.During use under field conditions (factors), under the situation that occurring in nature physical condition (temperature, moisture, the gas permeability of soil, sunlight), electrochemical conditions (soil mineral content and pH value), biotic condition (soil organic matter content and microbial population) all possess, repeatability reaches 80%, as long as keeping the advantage of these probioticss can use, as isolate the single variety bacterial classification, do not guarantee the non-leguminous plant nodulation and nitrogen fixation.
The science thinking of strain improvement and 5 step method are as follows:
(1) preparation of bacterial classification
The sublate method of sterile fermentation will according to symbiotic relationship and the intergrowth relation of microorganism, mutually according to mechanism, be carried out integration of science by optimization principles from gathering the strong bacterial classification of biological activity of coming naturally.Based on Rhizobium leguminosarum, be aided with the endophyte in the non-leguminous plant root system, form the nitrogen-fixing microorganism population, the mixing of making up bacterial classification is drawn and is grown;
(2) substratum is according to the demand of microorganism at the occurring in nature life condition, adopts available natural medium under the natural condition, possesses competent carbon source and nitrogenous source to guarantee fermenting process, and the people is for being aided with the needed various trace elements of microorganism growth, as; Iron (Fe), calcium (Ca), sodium (Na), potassium (K), magnesium (Mg), molybdenum (Mo) etc.
(3) open fermentation
For guaranteeing the powerful vitality of flora, must ferment under field conditions (factors) (flora propagation, expansion), but fermenting process is avoided living contaminants most, under physical environment, prevents from again to pollute again, for this reason, adopt the method that strengthens inoculum size, caused the bacterial classification population advantage at fermentation initial stage, made full use of the antagonism relation of microorganism, dominance principle, that guarantees to ferment normally carries out;
(4) tieback of bacterial classification and detection
After the fermentation, the flora that propagation is enlarged applies just at the growing plants root system, will use this moment in multiple pulse family and non-leguminous plant, observe root nodule and the root nodule situation tied, gather nodule detection again and have or not nitrogenase activity, separate endophyte kind kind and quantity and the bacterial classification that connects and compare, when kind, quantity near the time, the nodulation and nitrogen fixation effect is arranged again, and selecting the strongest active flora is bacterial classification of the present invention;
(5) be meant animal, plant, microorganism growth optimum condition under the so-called natural condition, sufficient nutrition (carbon-nitrogen ratio), 24 ℃-34 ℃ of temperature, humidity 30%-80%, pH value 6-8, well-ventilated, weather condition such as sufficient oxygen, nitrogen had both been guaranteed, the well-ventilated also is not the air feed abundance, relevant with gaseous interchange between plant, soil, space, the solar radiation abundance.Above condition be to guarantee that bacterial classification is effectively requisite, and this is difficult to guarantee in the laboratory.
Open fermentation is produced liquid fertilizer.Spontaneous induction non-leguminous plant nodulation and nitrogen fixation technology is directly used in agriculture production and produces bio-feritlizer exactly, can extend the high-performance solid bio-feritlizer that is used for making the agriculture production broad practice, these bio-feritlizer characteristics are the fabaceous root nodule bacterium of gathering from occurring in nature, " endophyte " in the non-leguminous plant root system, vinelandii in the soil, and actinomycetes, fungi, carry out scientific combination, optimize screening, the nature rejuvenation, fermentative production comes out approaching to carry out under the natural condition as far as possible with natural medium.Enlarge bacterial classification with the open fermentation method, this bacterium liquid is exactly liquid fertilizer, has broad spectrum, can be used for pulse family and and non-leguminous plant, comprise field crop, woods fruit, vegetables, flowers, because can open fermentation, suitability for industrialized production easily.
Can adopt the intermittent type liquid fermenting to produce liquid fertilizer or continuity solid fermentation production solid fertilizer:
The intermittent type liquid fermenting, that is: fermentation for some time stops, and adds the plural process that then ferments again behind the novel bacterial.
The continuity solid fermentation that is: behind the first flipover solid fermentation, carries out the fermentation of accumulation formula again after the pulverizing, carry out the process of static fermentation after pulverizing again again.
Mass production bacterium liquid, key issue are the degenerate problems of bacterial classification, aborning, and the tieback and the detection of the seed selection science thinking of this bacterial classification of the present invention and 5 step method, particularly bacterial classification repeatedly, otherwise can not guarantee the quality of liquid fertilizer.
The solid fermentation bio-feritlizer.The present invention is based on breeding species under the natural condition, open fermentation need not medium sterilization, so one group of seed selection is fit to the bacterial classification of farm crop, open fermentation is expanded into the secondary bacterial classification, and inoculating in the solid medium repeatedly ferments through solid can produce solid fertilizer.This fertilizer using method is suitable, and natural condition are good.Can make the non-leguminous plant nodulation and nitrogen fixation.Even temporary transient non-nodulating also has in the cell of flora invaded plants root system, reach nitrogen fixation.Make farm output increase quality improving.Can think that this is a kind of brand-new environmental friend fertilizer.
The definition of bio-feritlizer of the present invention: bio-feritlizer is the fabaceous root nodule bacterium of gathering from occurring in nature, and " endophyte " in the non-leguminous plant root system carries out scientific combination; Optimize screening, natural rejuvenation, fermentative production comes out approaching to carry out under the natural condition as far as possible with natural medium.
The principal feature of bio-feritlizer:
Broken the boundary of leguminous plants and non-leguminous plant, both can on leguminous plants, use, also can on non-leguminous plant, use: both can make Inoculant and use, also can use as base manure; Both can use, also can use in the extracting solution mode in the solid mode; Both can be used in the root of farm crop, also can spray on the cauline leaf of farm crop;
This is because the activity of existing vitality intensive microorganism in the bio-fertilizer, also comprises because of materials very useful to farm crop such as polybasic growth hormone that these useful populations produce in its complicated vital movement process, regulin, enzyme, protein, amino acid:
Bio-feritlizer has developed several generations, gets for the 5th generation now, and overtesting is all done in each province and city, area except that Tibet in the whole nation, and the experimental plot on up to ten million mu of ground and the trial report of enormous amount are arranged.Mass production facts have proved that this fertilizer has obvious social and economic benefit.
This fertilizer is applied to many non-leguminous plants such as ginger, tomato, strawberry, Elizabeth melon and has all reached the good effect of nodulation and nitrogen fixation crop quality.Laiwu City Gao Zhuan town, Shandong Province one peasant uses this fertilizer plantation ginger, the output of ginger is enhanced about more than once, prevented ginger plague again, the quality of ginger obviously improves, and through chemical examination, the chemical examination unit is Inst. of Plant Nutrition ﹠. Resource, Beijing City Academy of Agricultural ﹠. Fo the ginger of using this fertilizer and not using this fertilizer, the chemical examination time is on December 4th, 2002, and result of laboratory test sees Table 1.
Table 1
| The check numbering | The sample title | Test item and result | ||||||
| Crude protein % | Coarse ash % | Vc Mg/kg | Soluble sugar mg/kg | Solubility acid mg/kg | Total amino acid content % | Nitric nitrogen mg/kg | ||
| 02287-1 | Fresh ginger (contrast) | 1.74 | 0.98 | 7.44 | 3.45 | 0.128 | 0.382 | 370 |
| 02287-2 | Fresh ginger (G formula bio-fertilizer) | 2.43 | 0.87 | 8.31 | 2.35 | 0.117 | 0.288 | 266 |
| Following space | ||||||||
Chinese Academy Of Preventive Medicine Research Institute Of Nutrition And Food Hygiene is using this fertilizer and the tomato of not using this fertilizer, strawberry, the chemical examination of Elizabeth melon process.Use tomato, the strawberry of this fertilizer, contained composition carotene, the vitamins C to the human body beneficial of Elizabeth melon, qualities such as sugar, acids improve greatly.See Table 2, table 3, table 4; Table 2 is that tomato is used this fertilizer and the test report table of not using this fertilizer; Table 3 is that strawberry is used this fertilizer and the test report table of not using this fertilizer, and table 4 is that the Elizabeth melon is used this fertilizer and the test report table of not using this fertilizer.
Table 2
Chinese Academy Of Preventive Medicine Research Institute Of Nutrition And Food Hygiene's test report
| Interventions Requested and result | ||||||||
| The sample title | Carotene (μ g/100g) | Vitamins C (mg/100) | Total reducing sugar (g/100g) | Total acid (g/kg) is with citrometer | Phosphorus (mg/100g) | Potassium (mg/100g) | Calcium (mg/100g) | Zinc (mg/100g) |
| Tomato (bio-feritlizer) | 456.0 | 16.2 | 3.6 | 4.74 | 36.0 | 134.4 | 7.83 | 0.13 |
| Tomato (contrast) | 379.0 | 10.2 | 1.4 | 4.27 | 43.0 | 117.6 | 15.4 | 0.15 |
Table 3
Chinese Academy Of Preventive Medicine Research Institute Of Nutrition And Food Hygiene's test report
| Interventions Requested and result | ||||
| The sample title | Carotene (μ g/100g) | Vitamins C (mg/100) | Total reducing sugar (g/100g) | Total acid (g/100g) is with citrometer |
| Strawberry (bio-feritlizer) | 6.4 | 71.98 | 5.6 | 6.0 |
| Strawberry (contrast) | Do not detect | 30.23 | 4.3 | 6.6 |
Table 4
Chinese Academy Of Preventive Medicine Research Institute Of Nutrition And Food Hygiene's test report
| Bacterial manure group (mg/100g | Control group (mg/100g) | The bacterial manure group | Control group | ||
| Aspartic acid | 128.3 | 101.2 | Moisture | 86.6g/100g | 88.3g/100g |
| Threonine | 20.4 | 17.6 | Ash content | 0.58g/100g | 0.58g/100g |
| Serine | 43.5 | 38.4 | Protein | 1.16g/100g | 0.97g/100g |
| L-glutamic acid | 514.5 | 422.5 | Total reducing sugar | 12.9g/100g | 11.5g/100g |
| Glycine | 31.3 | 24.7 | Total acid | 1.716g/100g | 2.034g/100g |
| L-Ala | 46.1 | 60.3 | Carotene | 59.6μg/100g | 39.9μg/100g |
| Xie Ansuan | 28.8 | 21.6 | Food fibre | 0.88g/100g | 0.62g/100g |
| Methionine(Met) | 16.3 | 11.0 | VITMAIN B1 | 0.04g/100g | 0.03g/100g |
| Isoleucine | 20.2 | 15.1 | Wei ShengsuB2 | 0.026g/100g | 0.023g/100g |
| Leucine | 30.4 | 24.6 | Vitamins C | 8.33g/100g | 6.07g/100g |
| Tyrosine | 10.6 | 8.5 | Nicotinic acid | 1.00g/100g | 0.73g/100g |
| Phenylalanine | 19.1 | 16.3 | Carbohydrate | 10.78g/100g | 9.53g/100g |
| Histidine | 11.6 | 9.2 | Heat | 47.8 kilocalorie/100g | 42.0 kilocalorie/100g |
| Methionin | 25.6 | 21.4 | |||
| Arginine | 36.5 | 33.0 | |||
| Proline(Pro) | 17.6 | 15.7 | |||
The definition of bio-feritlizer is: the bio-feritlizer definition of producing based on the technology of the present invention is because biological vital movement can directly or indirectly provide the nutrient of crop normal development growth, and can improve soil, improves the material of the yield and quality of crop.Bio-feritlizer is different from fertilizer, inorganic fertilizer, chemical fertilizer, but should contain lid " bacterial manure " " microbial inoculum " " Inoculant ", life conditioning agent, growth hormone ... etc.
Technology of the present invention, because powerful microorganism species and the open fermentation technology useful to agricultural are arranged, so energy
Be used for the processing of domestic waste and industrial organic waste materials.Promptly these organism are produced bio-feritlizer as solid medium, can be through solid fermentation repeatedly, the bio-feritlizer of production various uses and different class.This has stack the naturally fermentation and the aerobic fermentation of rubbish to produce compost.But adding bacterial classification of the present invention in each degradation period of rubbish, final production goes out efficiently, the bio-feritlizer of high value.
Claims (1)
1, the bacterial classification of spontaneous induction non-leguminous plant nodulation and nitrogen fixation, its sample is preserved at China Committee for Culture Collection of Microorganisms common micro-organisms center, the bacterial classification that it is characterized in that described non-leguminous plant nodulation and nitrogen fixation is the combination flora, registers on the books to number and be in the bacterial classification of combination flora and preservation center thereof:
Register on the books and number 0949 in viride (Trichoderma viride) preservation center
Register on the books and number 0948 in root nodule bacterium (Rhizobium sp.) preservation center
Register on the books and number 0947 in look brown streptomycete (Streptomyces chromofuscus) preservation center
Register on the books and number 0946 in Bacillus foecalis alkaligenes (Alcaligenes faecalis) preservation center
Register on the books and number 0945 in edaphic bacillus (Agrobacterium sp.) preservation center
Register on the books and number 0944 in bacillus megaterium (Bacillus megateriium) preservation center
More than these bacterial classifications must combine, in 0 ℃ of preservation of low temperature, repeatability reaches 80%, as long as keep the advantage of these probioticss to use, as isolates the single variety bacterial classification, does not guarantee the non-leguminous plant nodulation and nitrogen fixation.
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| CN102747002B (en) * | 2011-12-27 | 2013-09-18 | 陕西省能源化工研究院 | Agrobacteriumsp. having free-living nitrogen fixing ability, and applications thereof |
| CN103695320B (en) * | 2013-12-26 | 2015-11-18 | 杨文慧 | Microorganism and preparing the application in bio-feritlizer, bio-feritlizer and application thereof |
| CN105130652A (en) * | 2015-09-09 | 2015-12-09 | 张乔木 | Organic composite water soluble fertilizer and preparation method thereof |
| CN106134728A (en) * | 2016-07-04 | 2016-11-23 | 东莞市保得生物工程有限公司 | A kind of method utilizing root nodule bacteria to promote Non-legume plants growth |
| CN110115212A (en) * | 2019-06-27 | 2019-08-13 | 安徽新熙盟生物科技有限公司 | A kind of implantation methods using rhizobium in Non-legume plants |
| CN113373074A (en) * | 2020-02-24 | 2021-09-10 | 刘�文 | Method for nodulation and nitrogen fixation of non-leguminous crops and method for cultivating microbial inoculum, preparation and application |
| CN113005064B (en) * | 2021-03-29 | 2022-07-26 | 西北农林科技大学 | Alcaligenes and application thereof and method for promoting soybean growth |
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